CN113995052A - Grassiness sugarcane tail leaf silage and preparation method thereof - Google Patents
Grassiness sugarcane tail leaf silage and preparation method thereof Download PDFInfo
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- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention relates to the technical field of silage fermentation, in particular to elephant grass and sugarcane tail leaf silage and a preparation method thereof, and the best mass ratio of elephant grass to sugarcane tail leaf silage is obtained through the silage research of elephant grass and sugarcane tail leaf with different mass ratios; further experiments are carried out on the strains screened by the personnel of the project group to obtain the lactobacillus capable of effectively promoting the decomposition of the grassiness and sugarcane tail leaf silage; the plant extract is used for replacing a conventional accelerant, and the plant extract mixed accelerant suitable for growth of the dominant strain xf 09# is screened out, so that the growth of the strain xf 09# can be effectively promoted, the fermentation efficiency is improved, the accelerant raw material used in the method is easy to obtain, the cost is low, the preparation method is simple, the accelerant is easy to obtain, and the accelerant is very friendly to self use of farmers.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of silage fermentation, in particular to grassiness sugarcane tail leaf silage and a preparation method thereof.
[ background of the invention ]
The elephant grass is a high-quality green feed for ruminants in south China, and is a raw material for preparing silage in Guangxi at present, because the elephant grass has high biological yield, good palatability and high nutritional value and can grow under different soil types, fertility levels and weather conditions to be planted in large quantities; the sugarcane tail leaves are main byproducts of the harvested sugarcane, and have the advantages of large yield, high sugar content and good palatability. Comprehensively considering conventional nutrient components, in-vitro fermentation characteristics, rumen degradation rule and the like, the sugarcane tail leaves are coarse feed resources with high nutritional values and are required to be developed and utilized; the existing research shows that the silage of the sugarcane tail leaves can achieve good fermentation effect within 30 days, and the silage is an effective storage mode of the sugarcane tail leaves. The prepared silage not only can improve the forage grass suitabilityIt is orally administered and can increase animal feed intake. But for some pastures with the characteristics of high moisture content, low soluble carbohydrate (WSC) and the like, the quality of the single silage is poor, and the mixed silage can improve the WSC content, inhibit poor fermentation and improve the fermentation quality of the silage; in silage, as the ratio of tail leaves of sugarcane increases, the dry matter recovery rate and the lactic acid content of the silage are remarkably increased, and the pH value, acetic acid and NH are increased3N pole is obviously reduced, and the quality of the water hyacinth silage can be obviously improved by the sugarcane tail leaves. The sugarcane tail leaves are rich and cheap feed resources in Guangxi regions, so that the research on the silage mixed with the sugarcane tail leaves and the elephant grass is particularly important; can provide theoretical reference for preparing high-quality grassiness silage and efficiently utilizing the sugarcane tail leaf coarse fodder resource.
Adding lactic acid bacteria to silage is a commonly used technical means at present for silage, and the applicant subject group also aims to screen more dominant strains for silage processing, for example, the applicant patents: 201911373265.5 discloses 4 dominant lactic acid bacteria, and long-term research on the strains shows that the 4 dominant lactic acid bacteria can effectively promote the decomposition of the feed when applied to silage, but practical experiments show that: different lactobacillus strains have different fermentation effects on different silage substrates, and not all the lactobacillus strains can promote the silage fermentation of different substrates, so that experiments are needed to research if lactobacillus is applied to different silage; in order to effectively utilize two raw materials of grassiness and sugarcane tail leaves to prepare the silage, the fermentation conditions need to be continuously optimized.
[ summary of the invention ]
In view of the above, in order to effectively utilize two raw materials, i.e., grassiness and sugarcane tail leaves to prepare the silage, the fermentation conditions need to be continuously optimized, and the aims of improving the disappearance rate of dry substances outside the silage body and improving the quality of the silage are achieved mainly by adjusting the mixing ratio of the grassiness and sugarcane tail leaf raw materials, optimizing strains suitable for the fermentation of the grassiness and sugarcane tail leaf raw materials and optimizing promoters suitable for the fermentation strains.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the pennisetum purpureum leaf and sugarcane tail leaf silage is prepared by mixing and ensiling pennisetum purpureum and sugarcane tail leaves according to the mass ratio of 1: 3.
Further, Lactobacillus fermentum xf 09# strain is added into the silage feed according to the inoculation amount of 0.1% in the silage process; the preservation number of the lactobacillus fermentum is CCTCC NO. M2019639.
Further, the promoter of Lactobacillus fermentum xf 09# strain is: folium Eucalypti Globueli extractive solution, folium Mori extractive solution and/or radix Puerariae extractive solution.
Further, the promoter of Lactobacillus fermentum xf 09# strain is: the eucalyptus leaf extract, the mulberry leaf extract and the kudzu leaf extract are as follows by mass: 1-2:2-4: 1-3.
Further, the promoter of Lactobacillus fermentum xf 09# strain is: the eucalyptus leaf extract, the mulberry leaf extract and the kudzu leaf extract are as follows by mass: mixing at ratio of 1:4: 3.
Further, the preparation method of the eucalyptus leaf extracting solution comprises the following steps: mixing fresh folium Eucalypti Globueli and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 30min, filtering, and diluting the filtrate by 10 times;
the preparation method of the mulberry leaf extracting solution comprises the following steps: mixing fresh folium Mori and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times;
the preparation method of the kudzu leaf extracting solution comprises the following steps: mixing fresh folium Puerariae Lobatae and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times.
The invention also comprises a method for preparing the grassiness sugarcane tail leaf silage, which comprises the following steps: crushing elephant grass and sugarcane, mixing according to the fresh weight ratio of 1:3, and adding an accelerator into a substrate according to the addition amount of 10 g/kg; after 2h of stack retting, Lactobacillus (Lactobacillus fermentum) xf 09# strain is inoculated according to the inoculum size of 0.1%, the mixture is evenly mixed, sealed by a vacuum packaging machine and then placed in a dark place for fermentation for 45 days at normal temperature.
Further, the accelerant is prepared by mixing a mixed solution of a eucalyptus leaf extracting solution, a kudzu leaf extracting solution and a mulberry leaf extracting solution according to the mass ratio of 1:4: 3.
The invention also comprises the application of the grassiness sugarcane tail leaf silage in improving the dry matter disappearance rate of the feed silage.
The inoculation referred to in this application is an inoculum solution, i.e.: mixing the bacterial powder and distilled water according to a solid-to-liquid ratio of 1:400 to obtain stock strain liquid with a viable count of about 106cfu/mL。
The invention has the following beneficial effects:
1. the invention obtains the best mass ratio of the grassiness to the sugarcane tail leaf silage through the silage research of the grassiness and the sugarcane tail leaf with different mass ratios; further experiments are carried out on the strains screened by the personnel of the project group to obtain the lactobacillus capable of effectively promoting the decomposition of the grassiness and sugarcane tail leaf silage; the plant extract is used for replacing a conventional accelerant, and the plant extract mixed accelerant suitable for growth of the dominant strain xf 09# is screened out, so that the growth of the strain xf 09# can be effectively promoted, the fermentation efficiency is improved, the accelerant raw material used in the method is easy to obtain, the cost is low, the preparation method is simple, the accelerant is easy to obtain, and the accelerant is very friendly to self use of farmers.
[ description of the drawings ]
FIG. 1 is a graph of growth curves of different extracts on strain xf 09 #.
[ detailed description ] embodiments
All of the features disclosed in this specification, or all of the steps in any method or process so disclosed, may be combined in any combination, except combinations of features and/or steps that are mutually exclusive.
Any feature disclosed in this specification (including any accompanying claims, abstract) is merely an example of a generic series of equivalent or similar features, unless explicitly described as such.
Example 1:
the embodiment is a preparation method of elephant grass and sugarcane tail leaf mixed silage.
In the embodiment, the raw materials are the grassiness and the sugarcane tail leaves without any other additives, and the influence of different mixing ratios on the ensiling fermentation quality and the in vitro dry matter disappearance rate of the grassiness and the sugarcane tail leaves is mainly researched as follows:
1. test materials: the grassiness and the tail leaves of the sugarcanes used in the test are respectively taken from a grassy planting base of the Guangxi buffalo research institute and a sugarcane planting field in the Yangxi Nanning city, Yongxi province and county. Cutting the two mixed storage raw materials to about 2cm for later use. The conventional nutritional ingredients of the mixed stock are shown in table 1:
TABLE 1 nutritional ingredients of elephant grass and sugarcane tail leaves before ensiling
2. The test method comprises the following steps: weighing the grassiness and the sugarcane according to the fresh weight ratio of 100:0, 75:25, 50:50, 25:75 and 0:100 (groups I, II, III, IV and V) by adopting a small-scale fermentation method, uniformly mixing, carrying out 5 treatments, repeating 5 treatments, filling each treatment into polyethylene film bags with 500g each bag, sealing by using a vacuum packaging machine, fermenting at normal temperature in a dark place for 45 days, and then unsealing, sampling and analyzing;
3. the measuring index and the measuring method of the silage comprise the following steps:
fermentation quality: accurately weighing 20g of sample per bag after opening the seal, putting the sample into a 250mL wide-mouth bottle, adding 180mL of distilled water, sealing the bottle cap by using a sealing film, putting the bottle cap into a refrigerator for leaching at 4 ℃ for 24 hours, then filtering by using 2 layers of gauze, and measuring the pH value and the content of organic acids (lactic acid, acetic acid, propionic acid and butyric acid) by using the filtered silage leaching liquor. The pH of the sample was measured using a portable pH meter. Centrifuging the leaching solution at 2000r/min for 5min, filtering with 0.45 μ M filter membrane, and measuring organic acid content with high performance liquid chromatography (Agilent 1260) (chromatographic column KC-811, detector SPD-M10AVP, column temperature 50 deg.C, flow rate 1mL/min, detection wavelength 210nm, and sample introduction amount 5 μ L). NH3-N was measured using a phenol-sodium hypochlorite colorimetric method.
② nutrient components: weighing 200g of sample per bag after opening and sealing, placing in a drying oven at 65 ℃ for drying to constant weight, recording the weight, crushing, measuring the content of bound water, and calculating the content of Dry Matter (DM) of the sample; measuring the content of Crude Protein (CP) by adopting a Kjeldahl method; the Organic Matter (OM) content was determined by 920.39 method in AOAC; NDF and acid-scoured fiber (ADF) content were determined by Van Soest et al.
③ in vitro fermentation characteristics: experimental rumen fluid donor a 3-head body weight of a stock farm planted in the research institute for buffalo, guangxi was about (65050kg) a female buffalo fitted with a permanent fistula. The daily ration is a daily feed formula of Guangxi buffalo institute, and free drinking water is fed twice (08:00 and 15:00) every day. Placing rumen fluid in a vacuum flask before morning feeding, rapidly returning to laboratory, mixing in equal volume, filtering with 4 layers of gauze to a preheated collection bottle, introducing CO2Maintaining an anaerobic environment. Mixing buffer solution (pH 6.80) prepared by McDougll's method with rumen fluid at volume ratio of 4:1 to obtain culture solution, injecting 50mL of culture solution into culture bottle containing 0.5g of sample, introducing CO2Discharging air and keeping anaerobic environment, immediately covering a rubber plug and an aluminum cover, compacting by using a special sealing clamp, culturing for 48 hours at a constant temperature of 39 ℃ under constant vibration (100r/min), and then measuring the gas production, the pH value, the Volatile Fatty Acid (VFA) and the in vitro dry matter disappearance rate, wherein the VFA content is measured by using gas chromatography (Agilent 7890A).
Data sorting and analyzing: after the basic data are arranged by Excel software, single-factor variance analysis is carried out by SAS 9.2 statistical software, and multiple comparison of the data is carried out by a Tukey method, wherein the significance level of P is less than 0.05. The results obtained from the tests are shown in table 2:
TABLE 2 influence of different mixing ratios on the fermentation quality of mixed silage of grasses and sugarcane tail leaves
Note: the labeling of different lower case letters in the same column indicates significant difference (P < 0.05); the following table shows that the ratio of the grassiness to the tail leaves of the sugarcanes is 100:0, the ratio of the grassiness to the tail leaves of the sugarcanes is 75:25, the ratio of the grassiness to the tail leaves of the sugarcanes is 50:50, the ratio of the grassiness to the tail leaves of the sugarcanes is 25:75, the ratio of the grassiness to the tail leaves of the sugarcanes is 0:100, and ND shows that the grassiness and the tail leaves of the sugarcanes are not detected.
As can be seen from Table 2, the pH values of the mixed fodder in groups II, III, IV and V are significantly lower than that in group I (P)<0.05), and groups IV and V are significantly lower than groups II and III (P)<0.05). The lactic acid content in group IV and group V is significantly higher than that in group I, II and III (P)<0.05). Group V acetic acid content is significantly lower than groups I, II, III and IV (P)<0.05). No propionic acid was detected in any of the mixed feeds. No butyric acid is detected after a certain proportion of the sugarcane tail leaves are added into the mixed storage feed of the elephant grass and the sugarcane tail leaves. NH (NH)3The content of N in group I is significantly higher than that in groups II, III, IV and V (P)<0.05)。
The results of the effect of different mixing ratios on the nutrient content of the mixed silage of grasses and sugarcane tail leaves are shown in Table 3.
TABLE 3 Effect of different mixing ratios on the Mixed silage Nutrition ingredients of pennisetum sinese L and sugarcane Tail leaves
As can be seen from Table 3, the DM content VIXIII group was significantly different between the groups (P < 0.05). CP content IIIIIIVV groups, and significant differences (P <0.05) exist among the groups. OM content VIRIII I, where IV and V are significantly higher than I, II and III (P < 0.05). The NDF content VIVIII II I, where V is significantly higher than I (P < 0.05). ADF content of mixed storage feed was not significantly different among groups (P > 0.05).
The influence of different mixing ratios on the in vitro fermentation parameters of the mixed silage of grasses and sugarcane tail leaves is shown in table 4:
TABLE 4 influence of different mixing ratios on in vitro fermentation parameters of mixed silage of grasses and sugarcane tail leaves
As can be seen from Table 4, the in vitro dry matter disappearance of the compost increased with the increase of the ratio of the tail leaves of sugarcane, with the highest being group IV (group IV V III II I), but with insignificant difference between the groups (P > 0.05). The gas production rates IV, III, II, and V, wherein IV is significantly higher than V, III, II, and I (P <0.05), and III is significantly higher than I and V (P < 0.05). The pH values of groups I and II are significantly lower than those of groups III, IV and V (P < 0.05). Total VFA contents II, III and V were significantly higher than in group I (P < 0.05). The acetic acid content in groups II, III and V is significantly higher than in group I (P < 0.05). The propionic acid content in groups II and III is significantly higher than in group I (P < 0.05). Butyric acid contents of groups II and III were significantly higher than those of group I (P < 0.05). The NH3-N content was minimal for group V and significantly lower than for group III (P < 0.05).
The results show that the pH values of the mixed storage feed are obviously lower in groups II, III, IV and V than in group I, and in addition, the groups IV and V are obviously lower than in groups II and III; the lactic acid content in groups IV and V is obviously higher than that in groups I, II and III; group V, the acetic acid content of which is significantly lower than that of groups I, II, III and IV; no butyric acid is detected after a certain proportion of the sugarcane tail leaves are added into the mixed storage feed of the elephant grass and the sugarcane tail leaves; NH (NH)3The N content in group I is significantly higher than in groups II, III, IV and V. This result shows that as the proportion of the tail leaves of sugarcane increases, the pH value of the compost, acetic acid and NH3The content of N is in a tendency of decreasing, and the lactic acid is in a tendency of increasing, probably because when the elephant grass and the sugarcane tail leaf which are mixed and mixed in proportion are mixed and ensiled, the increase of the proportion of the sugarcane tail leaf increases the content of soluble sugar, provides favorable conditions for the growth and the propagation of lactic acid bacteria, promotes the lactic acid fermentation, reduces the pH value, inhibits the growth of harmful microorganisms, reduces the consumption of other mixed bacteria on the nutrient of fermented feed, and increases the mixed content of the elephant grass and the sugarcane tail leafFermentation quality of the ensilage.
In the test, the contents of DM, OM, NDF and ADF are IV, III and II, namely in the mixed storage feed of grassiness and sugarcane tail leaves, the contents of DM, OM, NDF and ADF of the mixed storage feed show an ascending trend along with the increase of the proportion of the sugarcane tail leaves, which shows that the increase of the proportion of the sugarcane tail leaves can improve the nutritive value of the mixed silage to a certain degree.
In the test, the in vitro dry matter disappearance rate and in vitro gas yield of the mixed storage material are improved along with the increase of the proportion of the sugarcane tail leaves, wherein the highest is IV group, and the fermentation quality and the nutritional value of the mixed silage are the best when the proportion of the elephant grass to the sugarcane tail leaves is 25: 75. The pH value has a decisive influence on the change of a plurality of parameters of rumen fermentation, and is one of the most important indexes for evaluating the rumen fermentation condition. The precondition for the degradation of the ration ingested by the animals in the rumen is that the pH remains within the normal range. VFA not only provides energy substances for animals but also participates in various metabolic activities, and VFA is one of indexes reflecting rumen fermentation level. In the test, the total VFA and acetic acid of the groups II, III and V are obviously higher than that of the group I, the propionic acid and butyric acid contents of the groups II and III are obviously higher than that of the group I, and the fermentation quality and the nutritive value of the silage are improved probably after a certain proportion of sugarcane tail leaves are added into grassiness, so that the in-vitro fermentation degree of the silage is improved.
In conclusion, under the test conditions, the mixed silage of the mixed grassiness sugarcane tail leaves can improve the quality of the grassiness silage, and improve the nutritional value and the in-vitro dry matter disappearance rate. Meanwhile, the mixed fermentation effect of the pennisetum purpureum and the sugarcane is optimal, wherein the fresh weight ratio of the pennisetum purpureum to the sugarcane is 25:75 (the reduction ratio is 1: 3).
Example 2:
the influence of the addition of lactic acid bacteria on the silage:
in actual research, the applicant found that: the fermentation effects of different lactic acid bacteria on silage are different, and the applicant utilizes several strains screened by the subject group to perform experiments, and the experiments specifically comprise the following steps: lactobacillus fermentum xf 09# accession number: CCTCC NO. M2019639, the strain is preserved in China center for type culture Collection, and the address is: wuhan university, the preservation date is 19.8.2019, and the preservation number of Lactobacillus mucosae (Lactobacillus mucosae) xf 13# is as follows: CCTCC NO. M2019640, the strain is preserved in China center for type culture Collection, and the address is: wuhan university, the preservation date is 19.8.2019, and the preservation number of Lactobacillus acidophilus (Lactobacillus acidophilus) xf 32 #: CCTCC NO. M2019641, the strain is preserved in China center for type culture Collection, and the address is as follows: wuhan university, the preservation date is 19.8.2019, and the preservation number of Lactobacillus plantarum (Lactobacillus plantarum) xf 42 #: CCTCC NO. M2019642, the strain is preserved in China center for type culture Collection, and the address is: wuhan university, the preservation date is 2019, 8 and 19 days (the specific screening methods and classification standards of the strains are disclosed in the patent: 201911373265.5 application).
The specific experimental method is as follows:
1. experimental materials: according to the embodiment 1, the fresh weight ratio of the elephant grass to the sugarcane is 25:75 (the reduction ratio is 1:3), the mixing fermentation effect is optimal, and the elephant grass and the sugarcane are mixed according to the mass ratio to prepare the silage;
2. lactic acid bacteria: the following 5 strains were selected: lactobacillus fermentum xf 09#, lactobacillus mucosae xf 13#, lactobacillus acidophilus xf 32#, lactobacillus plantarum xf 42#, lactobacillus commercially available (purchased from galyi bioengineering, ltd, kayokogaku, shandong);
3. the test method comprises the following steps: adopting a small-scale fermentation method, weighing and uniformly mixing the grassiness and the tail leaves of the sugarcane according to the fresh weight ratio of 25:75 (the reduction ratio is 1:3), and setting 6 treatments: total 5 treatments, treatment 1: adding a strain lactobacillus fermentum xf 09#, mixing with grassiness and sugarcane tail leaves for fermentation, and treating 2: adding lactobacillus mucosae xf 13# and grassiness and sugarcane tail leaves, mixing, fermenting and treating 3: adding lactobacillus acidophilus xf 32#, grass and sugarcane tail leaves, mixing, fermenting and treating the mixture 4: adding lactobacillus plantarum xf 42#, grassiness and sugarcane tail leaves, mixing, fermenting and treating 5: adding commercially available lactobacillus, grassiness and sugarcane tail leaves, and mixing and fermenting; and (3) CK group: without adding any lactic acid bacteria, the mixed fermentation is carried out only by using the mixed fermentation of the elephant grass and the sugarcane tail leaves with the fresh weight ratio of 25:75 (the reduction ratio is 1: 3). Repeating the treatment for 5 times, filling 500g of the mixture into polyethylene film bags, sealing the bags by using a vacuum packaging machine, keeping the bags away from light, fermenting the bags at normal temperature for 45 days, unsealing the bags, sampling and analyzing the bags;
in this example, the influence of the quality of the feed, the silage nutrient content and the silage fermentation parameters in vitro (see example 1 for the detection method of the relevant parameters) is mainly studied as follows:
TABLE 4 sensory evaluation of fermented feed
| Group of | Smell(s) | Color | Texture of | Rating |
| xf 09# | 24.5 | 19.5 | 9 | Excellence in |
| xf 13# | 23.9 | 19.1 | 9 | Excellence in |
| xf 32# | 21.8 | 18.2 | 9 | Excellence in |
| xf 42# | 23.4 | 17.3 | 9 | Excellence in |
| Commercially available lactic acid bacteria | 22.9 | 18.6 | 9 | Excellence in |
| CK | 22.6 | 18.5 | 9 | Excellence in |
The sensory evaluation was performed with reference to "silage quality evaluation standards (trial) published by 1996 of the ministry of agriculture in China, and is specifically shown in table 5:
TABLE 5 sensory evaluation criteria
| Item | Smell(s) | Color | Texture of |
| Excellence in | Sweet sour flavor comfort (18-25) | Bright yellow (14-20) | Loose, soft and non-sticky (8-10) |
| Good effect | Sour taste (9-17) | Brown yellow (8-13) | Middle (4-7) |
| In general | Sour of pungent wine (1-8) | Middle (1-7) | Slightly sticky (1-3) |
| Disadvantages and quality | Rotten taste of rotten flavor (0) | Dark brown (0) | Sticky clumps (0) |
As can be seen from tables 4 to 5, the sensory effect of the fermented feed was best when lactobacillus strain lactobacillus xf 09# was added, but the difference was not large as compared with the other group and the CK group, which indicates that the sensory effect of sugar cane caudal leaf was not large when lactobacillus was used for fermentation, and the sensory quality of silage was slightly improved by lactobacillus strain lactobacillus xf 09 #.
Effect of different lactic acid bacteria on the nutritional ingredients of silage
TABLE 6 Effect of different lactic acid bacteria on the nutritional composition of silage
Note: the same column is labeled with different lower case letters to indicate significant difference (P < 0.05).
As can be seen from table 6, the dry matter content xf 09# is significantly lower than the other treatment groups (P <0.05), the crude protein content is significantly higher than the other treatment groups (P < 0.05); the above results show that most of the lactic acid bacteria fermentation had little effect on the nutritional ingredients of the silage, while strain xf 09# had a greater effect on the nutritional ingredients of the mixed silage of elephant grass and sugarcane tail leaves than the other strains.
The influence of the in vitro fermentation parameters of the mixed silage of grass and sugarcane tail leaves of different lactobacillus subjects is shown in table 7:
TABLE 7 influence of different lactic acid bacteria on the in vitro fermentation parameters of mixed silage of grasses and sugarcane tail leaves
As can be seen from table 7, the in vitro dry matter disappearance rate, the gas production rate, the total VFA and the acetic acid content of the strain xf 09# are significantly higher than those of the other treatment groups, thereby demonstrating that the strain xf 09# can significantly improve the fermentation degree of the grassiness and sugarcane tail leaf silage, accelerate the decomposition of the grassiness and sugarcane tail leaf silage, and make the silage more easily absorbed by animals compared with other lactobacillus strains.
Several experiments from example 2 show that: the addition of lactic acid bacteria can accelerate the decomposition of the silage to a certain extent, but the decomposition capacities of different lactic acid bacteria are different, and it is also shown that not all strains can play a role in accelerating the decomposition of the silage: the strain xf 42# is inferior to the CK group in its ability to decompose silage; in actual research, different substrates have different decomposition capacities to different lactobacillus strains due to different substrate ratios and different silage raw materials, some strains have a promoting effect, some strains have no effect, and even have a certain inhibiting effect, because the effective components of different silage raw materials are completely different, the strains have different decomposition capacities to the different lactobacillus strains, and in the silage process of some plants, the growth of some lactobacillus strains is possibly inhibited, so that dominant bacteria cannot be formed, and finally the silage decomposition capacity is reduced. Therefore, whether the lactobacillus can effectively decompose the feed by applying the silage can be determined by continuous bacteria screening and experiments of researchers. Therefore, strain screening and application identification are particularly important.
Example 3:
effect of promoters on strain fermentation:
in actual ensiling fermentation, in order to promote fermentation, certain additives and active ingredients are generally added to promote the proliferation of beneficial bacteria, and beneficial ingredients such as trace elements and amino acids are generally added to achieve the purpose of promoting the growth of strains under conventional experiments, but the ingredients are single, complex compounding experiments are generally required to be carried out, substances with more than 10 different ingredients are generally required to be combined, and for actual operation, professional personnel are generally required to prepare the ingredients, and the manufacturing cost of the ingredients is high; to this end, the subject has focused on various alternatives, considering: some plant extract liquid obtained locally in the current area can be used for replacing chemical promoters with complex components, although some plant extract liquid has reports of promoting the growth of microorganisms in the prior art, the plant extract liquid has very complex components and many conditions of inhibiting the growth of microorganisms, so if the plant extract liquid is used as the promoters, experimental screening is required; the plants (pasture or waste) which are common in the area are selected in the embodiment: eucalyptus leaves, mulberry leaves, alfalfa leaves, kudzu and pine needles.
Preparing a corresponding plant extract:
extracting eucalyptus leaves: mixing fresh folium Eucalypti Globueli and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 30min, filtering, and diluting the filtrate by 10 times.
② mulberry leaf extract: mixing fresh folium Mori and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times.
③ alfalfa leaves extract: mixing fresh alfalfa leaves and water according to the solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times.
Extracting the kudzu leaf liquid: mixing fresh folium Puerariae Lobatae and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times.
Fifthly, pine needle extract: mixing sun-dried folium Pini and water at solid-liquid mass ratio of 1:3, boiling, decocting for 30min, filtering, and diluting the filtrate by 10 times.
Preparation of the corresponding medium:
and (3) CK group: MRS liquid culture medium;
treatment 1: adding 10g/L of eucalyptus leaf extract into MRS liquid culture medium;
and (3) treatment 2: adding 10g/L mulberry leaf extract into MRS liquid culture medium raw material;
and (3) treatment: adding 10g/L alfalfa leaf extracting solution into the raw material of the MRS liquid culture medium;
and (4) treatment: 10g/L of kudzu leaf extracting solution is added into the MRS liquid culture medium raw material;
and (4) treatment 5: adding 10g/L pine needle extract into MRS liquid culture medium;
each treatment was repeated 3 times, and the group was inoculated with 3% of the inoculum size using the dominant strain of example 2: xf 09#, anaerobic culture at 37 ℃ and 1 time of bacterial liquid dilution with the same times every 3h, then measuring absorbance at 600nm, and drawing a growth curve, wherein the results are shown in figure 1: the lactic acid bacteria of the treatment group and the CK group both grew logarithmically, and had reached a stationary phase at 12h, treatment 1, treatment 2, and treatment 4 promoted the growth of the strain xf 09#, whereas treatment 3 and treatment 5 did not promote the growth of the strain xf 09#, and the growth rate was similar to that of the CK group, and treatment 4 was slightly lower than that of the CK group, indicating that the kudzu leaf extract of treatment 4 had an inhibitory effect on the strain xf 09 #.
To this end, the applicant selected treatment 1, treatment 2 and treatment 4 as accelerators, and carried out experimental selection of additive ratios using the following method: adding 10g/L accelerator mixed liquor into the MRS liquid culture medium raw material; the mixed solution is composed of eucalyptus leaf extract, kudzu leaf extract and mulberry leaf extract according to different percentages, then the strain is inoculated according to the inoculation amount of 0.1%, after anaerobic culture is carried out for 12 hours at the temperature of 37 ℃, the OD value is measured after the bacterial solution is diluted:
TABLE 8 levels of orthogonal Experimental factors for the growth Capacity of Strain xf 09#
The results of the orthogonal analysis of the above formulation are shown in Table 9:
TABLE 9 Quadrature experiments and results
| Number of experiments | Eucalyptus leaf extract | Kudzu leaf extract | Folium Mori extractive solution | OD600 |
| 1 | A | A | A | 3.14 |
| 2 | A | B | B | 3.05 |
| 3 | A | C | C | 3.31 |
| 4 | B | A | B | 2.64 |
| 5 | B | B | C | 2.83 |
| 6 | B | C | A | 2.74 |
| 7 | C | A | C | 2.31 |
| 8 | C | B | A | 2.42 |
| 9 | C | C | B | 2.46 |
| |
0 | 0 | 0 | 2.43 |
As is clear from Table 9, when 3 extracts were mixed and used, the amount of the eucalyptus leaf extract added was too large to inhibit the growth of the strain, even the CK group was equivalent, showing that: after the plant extract is mixed, the effective components in the plant extract may react, and the effective components do not necessarily have the effect of promoting the growth of the strains, so that the strain growth can be promoted by selecting the adding proportion of 1-7 groups in the extract mixed liquor, the strain growth effect is optimal and even superior to the growth promotion effect of a single extract by selecting the adding proportion of experiment 3, and for this reason, the promoter mixed liquor selects the following components in mass ratio: extracting a eucalyptus: kudzu leaf extracting solution: 1-2:2-4:1-3 of mulberry leaf extract; optimally selecting the adding proportion of experiment 3, namely the eucalyptus leaf extracting solution: kudzu leaf extracting solution: the ratio of the mulberry leaf extract to the mulberry leaf extract is 1:4: 3.
Example 4:
the experiment of example 3 shows that: mixing the plant extract according to the mass ratio: the eucalyptus leaf extracting solution: kudzu leaf extracting solution: the mulberry leaf extract can effectively promote the growth of lactic acid bacteria, and the application effect in the actual silage fermentation is not verified by experiments, so that the silage experiment is carried out by adopting the research result of the example 3, and the specific method is as follows:
experimental group 1: crushing elephant grass and sugarcane, mixing according to a fresh weight ratio of 25:75 (the reduction ratio is 1:3), and adding an accelerator according to the addition amount of 10 g/kg; wherein the accelerant comprises the following components in percentage by mass: the eucalyptus leaf extracting solution: kudzu leaf extracting solution: mixing the mulberry leaf extract with a mixed solution of 1:4: 3; after 2h of stack retting, inoculating strain xf 09# according to 0.1% of inoculum size, mixing uniformly, and setting 3 times of repetition; packaging into polyethylene film bags (500 g per bag), sealing with vacuum packaging machine, fermenting at room temperature in dark place for 45 days, and unsealing;
experimental group 2: crushing elephant grass and sugarcane, mixing according to a fresh weight ratio of 25:75, retting for 2h, inoculating the elephant grass and the sugarcane into a strain xf 09#, uniformly mixing, and repeating for 3 times; packaging into polyethylene film bags (500 g per bag), sealing with vacuum packaging machine, fermenting at room temperature in dark place for 45 days, and unsealing;
experimental group 3: crushing elephant grass and sugarcane, mixing according to a fresh weight ratio of 25:75 (the reduction ratio is 1:3), and adding an accelerator according to the addition amount of 10 g/kg; wherein the accelerant comprises the following components in percentage by mass: the eucalyptus leaf extracting solution: kudzu leaf extracting solution: mixing folium Mori extractive solution 1:4:3, mixing, and repeating for 3 times; packaging into polyethylene film bags (500 g per bag), sealing with vacuum packaging machine, fermenting at room temperature in dark place for 45 days, and unsealing;
control group: after smashing the elephant grass and the sugarcane, mixing according to the fresh weight ratio of 25:75, uniformly mixing, and repeating for 3 times; packaging into polyethylene film bags (500 g per bag), sealing with vacuum packaging machine, fermenting at room temperature in dark place for 45 days, and unsealing;
the silage in vitro fermentation parameters of the experimental group are tested, and the main measurement indexes are as follows: dry matter disappearance and total VFA in vitro, results are shown in table 10:
TABLE 10 in vitro fermentation parameters for differently treated mixed silage
| Test item | Experimental group 1 | Experimental group 2 | Experimental group 3 | Control group |
| In vitro dry matter disappearance (%) | 55.14a | 48.65b | 43.05c | 42.08c |
| Total VFA (mmol/l) | 25.31a | 21.15b | 20.18b | 19.24b |
Note: the marking of different lower case letters in the same row indicates that the difference is significant (P)<0.05)。
As can be seen from table 10, the in vitro dry matter disappearance and total VFA content of experimental group 1 were significantly higher than those of experimental groups 2-3 and the control group; the in vitro dry matter disappearance rate of the experimental group 2 is obviously higher than that of the experimental group 3 and the control group; the total VFA content is slightly higher than that of the experimental group 3 and the control group, but the difference is not obvious; the in vitro dry matter disappearance and total VFA content of experimental group 3 was not significantly different compared to the control group, thus demonstrating that: after the strain xf 09# is added, the fermentation of a mixture of sugarcane tail leaves and grassiness can be effectively promoted; the addition of the promoter can effectively promote the growth of the strain, improve the activity of the strain and further promote the fermentation of the strain xf 09# to silage; the fermentation efficiency of the experimental group 3 and the control group are close to each other, so that the inventors believe that the phenomenon has two reasons: firstly, the lactobacillus content is low in the ensiling process of elephant grass and sugarcane tail leaves, and the promoter cannot promote the growth of other microorganisms, so that the dominant lactobacillus is not formed, and the promotion effect is not obvious; ② the grassiness and sugarcane tail leaf silage process also contains other lactobacillus, however, the accelerant can not promote the growth of other lactobacillus strain, thus can not play the role of effectively improving and promoting silage fermentation. For whatever reason, the experiments of this example demonstrate that: the strain xf 09# can effectively promote the fermentation of sugarcane tail leaves and grassiness silage, and the growth of the strain xf 09# can be further promoted by adding the accelerant, so that the fermentation efficiency is improved.
To sum up, the application researches the silage fermentation of the mixing proportion of different grassiness and sugarcane tail leaves, and finally discovers that: when the grassiness: when the mass ratio of the sugarcane tail leaves is 25:75 (the reduction ratio is 1:3), the disappearance rate of dry matters in vitro is the highest, and the fermentation effect is the best; the applicant further researches on lactic acid bacteria screened by a subject group and finally discovers that the strain xf 09# can effectively promote the fermentation of mixed feed of grassiness and sugarcane tail leaves; through further research on the mixed solution of the plant extract, the plant extract additive is finally found to be capable of promoting the fermentation of the silage.
The above examples are merely illustrative of several embodiments of the present invention, and the description thereof is more specific and detailed, but not to be construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.
Claims (9)
1. The grassiness sugarcane tail leaf silage is characterized in that the silage is prepared by mixing and ensiling grassiness and sugarcane tail leaves according to the mass ratio of 1: 3.
2. The grassiness sugarcane tail leaf silage according to claim 1, characterized in that Lactobacillus fermentum (xf 09# strain) is added to the silage at an inoculation amount of 0.1%; the preservation number of the lactobacillus fermentum is CCTCC NO. M2019639.
3. The grassiness sugarcane tail leaf silage according to claim 2, characterized in that the promoter of Lactobacillus fermentum xf 09# strain is: folium Eucalypti Globueli extractive solution, folium Mori extractive solution and/or radix Puerariae extractive solution.
4. The grassiness sugarcane tail leaf silage according to claim 3, wherein the promoter of Lactobacillus fermentum xf 09# strain is: the eucalyptus leaf extract, the mulberry leaf extract and the kudzu leaf extract are as follows by mass: 1-2:2-4: 1-3.
5. The grassiness sugarcane tail leaf silage according to claim 3, wherein the promoter of Lactobacillus fermentum xf 09# strain is: the eucalyptus leaf extract, the mulberry leaf extract and the kudzu leaf extract are as follows by mass: mixing at ratio of 1:4: 3.
6. The grassy sugarcane tail silage according to any one of claims 3 to 5,
the preparation method of the eucalyptus leaf extracting solution comprises the following steps: mixing fresh folium Eucalypti Globueli and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 30min, filtering, and diluting the filtrate by 10 times;
the preparation method of the mulberry leaf extracting solution comprises the following steps: mixing fresh folium Mori and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times;
the preparation method of the kudzu leaf extracting solution comprises the following steps: mixing fresh folium Puerariae Lobatae and water at solid-liquid mass ratio of 1:3, heating to boil, decocting for 15min, filtering, and diluting the filtrate by 10 times.
7. A method of preparing a grassiness sugarcane tail silage as claimed in any one of claims 1-6, characterized in that the method is: crushing elephant grass and sugarcane, mixing according to the fresh weight ratio of 1:3, and adding an accelerator into a substrate according to the addition amount of 10 g/kg; after 2h of stack retting, Lactobacillus (Lactobacillus fermentum) xf 09# strain is inoculated according to the inoculum size of 0.1%, the mixture is evenly mixed, sealed by a vacuum packaging machine and then placed in a dark place for fermentation for 45 days at normal temperature.
8. The method as claimed in claim 7, wherein the promoter is prepared by mixing a mixture of eucalyptus leaf extract, kudzu leaf extract and mulberry leaf extract in a mass ratio of 1:4: 3.
9. Use of the grassiness sugarcane tail leaf silage according to any one of claims 1-6 for increasing the rate of dry matter disappearance from the feed silage body.
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